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Kolotnytska was born in 1972 in Brovary,[1][2] in the Ukrainian SSR of the Soviet Union (in present-day Ukraine). She graduated from an unspecified nursing college in Kiev and has been committed to her work since. Kolotnytska's husband, an electrician at a local factory, died in 1992, after which Kolotnytska moved from her local nursing job to a cook position at Ukraine's Antarctic research station. There she was described as "cooking marvelously" and "harshly enforcing hygiene rules".[1]

Kolotnytska moved to Libya in 2001, with the help of a medical agency acquired abroad, where she first worked at a hospital and then went on to serve Gaddafi.[2] At the time, Libya was a popular destination for Ukrainian physicians because the payments there were better than in Ukraine.[3] According to various estimates, there were about 500 Ukrainian doctors and nurses in Libya early 2011.[3] It is reported that Gaddafi employed four nurses from Ukraine.[4]

Kolotnytska was described by Gene Cretz, the U.S. ambassador to Libya, as the "voluptuous blonde" without whom Muammar Gaddafi, leader of Libya, never traveled.[10] The characterization surfaced after the secret diplomatic cable sent by Cretz was revealed by WikiLeaks as a part of its United States diplomatic cables leak. The cable was sent from the United States embassy in Tripoli on 29 September 2009. It was also revealed that the Libyan government sent a private jet to ferry her from Libya to Portugal to meet up with the Leader during his rest-stop. Kolotnytska alone is said to have "known Gaddafi's routine".[11][12] Kolotnytska's daughter Tatyana and her mother Iryna have categorically denied that Kolotnytska was the mistress of Gaddafi.[2][13]